TLR4通过下调ABCA1表达促进平滑肌源性泡沫细胞形成

发布时间：2018-01-21 00:01

本文关键词： 泡沫细胞血管平滑肌细胞 Toll样受体三磷酸腺苷-结合盒转运子A　出处：《第三军医大学学报》2014年07期 　论文类型：期刊论文

【摘要】：目的探讨Toll样受体4(Toll-like receptor 4,TLR4)促进平滑肌源性泡沫细胞的形成是否与三磷酸腺苷-结合盒转运子A1(ATP-binding cassette transporter,ABCA1)表达异常相关。方法组织贴块法培养原代血管平滑肌细胞(vascular smooth muscle cells,VSMCs);细胞分为野生型(Wild type)空白对照组、Wild type氧化低密度脂蛋白(oxLDL)组、TLR4敲除(TLR4-/-)空白对照组和TLR4-/-oxLDL组;油红O染色观察VSMCs内脂质聚集,并用异丙醇萃取油红O测定光密度值;RT-qPCR、Western blot检测VSMCs中TLR4、ABCA1、IL-1β及TNF-αmRNA和蛋白表达水平;细胞免疫荧光染色鉴定原代VSMCs及观察核转录因子-kappa B(NF-κB)核转位。结果①Wild type oxLDL组中见细胞内大量脂滴形成,油红O光密度值为Wild type空白对照组的2.3倍(P0.01);TLR4-/-oxLDL组中只见少许脂质沉积于细胞内,油红O光密度值与TLR4-/-空白对照组相比无明显变化(P0.05)。②Wild type oxLDL组ABCA1 mRNA及蛋白表达水平较Wild type空白对照组分别下降61%(P0.05),44%(P0.01);TLR4-/-oxLDL组ABCA1 mRNA及蛋白表达水平分别为TLR4-/-空白对照组的2.3倍(P0.05),1.7倍(P0.05)。③Wild type oxLDL组VSMCs中NF-κB核转位明显增多,IL-1β和TNF-αmRNA表达较Wild type空白对照组分别升高3.9倍(P0.05),1.7倍(P0.05);TLR4-/-oxLDL组VSMCs中无明显NF-κB核转位,仅IL-1βmRNA表达较TLR4-/-空白对照组升高1.2倍(P0.05)。结论 TLR4及其调节的炎性反应参与平滑肌源性泡沫细胞形成过程中ABCA1表达异常。
[Abstract]:Objective to investigate Toll-like receptor 4 (Toll-like receptor 4) of Toll like receptors. TLR4) promotes the formation of smooth muscle derived foam cells and ATP-binding cassette transporter with adenosine triphosphate binding cassette transporter A1. Methods the primary vascular smooth muscle cells were cultured by tissue patch method. The cells were divided into wild type (wild type) control group and wild type oxidized low density lipoprotein (LDL) group. TLR4 knockout TLR4 / -) blank control group and TLR4-/-oxLDL group; Oil red O staining was used to observe the accumulation of lipid in VSMCs and the optical density was determined by isopropanol extraction. The expression levels of IL-1 尾, TNF- 伪 mRNA and protein in VSMCs were detected by RT-qPCRG Western blot. Identification of Primary VSMCs by Cellular Immunofluorescence staining and observation of Nuclear transcription Factor -kappa Bon NF- 魏 B. Results 1 A large number of lipid droplets were found in the wild type oxLDL group. The optical density of oil red O was 2.3 times that of Wild type control group (P 0.01). In TLR4-/-oxLDL group, a little lipid was deposited in the cells. The optical density of oil red O was not significantly different from that of TLR4-r-blank control group (P0.05). 2 the expression of ABCA1 mRNA and protein in wild type oxLDL group decreased 61% and P 0.05 respectively compared with Wild type control group. P0.01C; The expression level of ABCA1 mRNA and protein in TLR4-/-oxLDL group was 2.3-fold of that in control group (P 0.05). The nuclear translocation of NF- 魏 B in VSMCs was significantly increased in the group of 1.7-fold P0.05N. 3 Wild type oxLDL. The expression of IL-1 尾 and TNF- 伪 mRNA was 3.9-fold higher than that of Wild type control group. No significant nuclear translocation of NF- 魏 B was observed in VSMCs of TLR4-/-oxLDL group. The expression of IL-1 尾 mRNA was only 1.2-fold higher than that of TLR4-r-blank control group (P0.05). Conclusion TLR4 and its inflammatory response are involved in the abnormal expression of ABCA1 in the formation of smooth muscle derived foam cells.
【作者单位】：第三军医大学大坪医院野战外科研究所神经内科;
【基金】：国家自然科学基金面上项目(81271282) 重庆市杰出青年科学基金(CSTC2012JJJQ10003)~~
【分类号】：R329.2
【正文快照】： 泡沫细胞聚集是动脉粥样硬化最主要的特征,在中晚期动脉粥样硬化病变中,巨噬细胞源性泡沫细胞约占30%,而平滑肌细胞源性泡沫细胞高达45%[1]。巨噬细胞源性泡沫细胞形成机制已有大量研究,而平滑肌细胞源性泡沫细胞形成机制的研究却很少。研究证实,在动脉粥样硬化斑块中,Toll样

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